Definition of a blood type in the two-dimensional way

A. Definition of a blood type consists in the two-dimensional way in simultaneous definition of group agglutinogens in erythrocytes of an blood on tsoliklona and group agglutinins in Serum of the studied blood by means of standard erythrocytes.

B. For definition of a blood type in the two-dimensional way except monoclonal antibodies anti-A, anti-B and anti-AB use standard erythrocytes of O(I), A (II) and B (III) group.

B. The blood for mixing is taken from a vein or a place of a nyxis of a finger in a dry pure test tube. The blood is centrifuged or left alone for 20-30 minutes for unit of Serum.

G. Definition is made on a white plate on which top part apply designations from left to right: anti-A, Anti-B. At the upper edge inscribe a surname and initials of the person at whom define a blood type.

D. Under the corresponding designations of blood types apply on a plate on one big drop (0,1 ml) of standard monoclonal antibodies.

E. Under designations of O(I), A (II) and B (III) apply on the right part of a plate on one small (0,01 ml) to a drop of standard erythrocytes in the following order from left to right: O (I), A(II) and B (III).

Zh. From the test tube containing the patient's blood, a pipette extract Serum and drop it on one big (0,1 ml) a drop on the prepared standard erythrocytes. After that the same pipette gather erythrocytes of an testing blood since a bottom of a test tube and put them on small (0,01 ml) to a drop near each drop of the prepared monoclonal antibodies.

H. In all drops of an antibody and Serum carefully mix with erythrocytes, using glass rods, the plate is shaken, then for 1-2 minutes left alone and again periodically shaken. Observation over the course of reaction is made by not less than five minutes.

I. In process of offensive of agglutination with standard erythrocytes, but not earlier than in 3 minutes, in those drops, in which it I came, add on one drop (0,05 ml) of isotonic NaCl solution and continue observation when rocking a plate before the efflux of five minutes.

The accounting of reaction is made by comparison of the results received by means of standard Serums and erythrocytes.

Results of the reactions received by means of monoclonal antibodies and standard erythrocytes have to coincide, i.e. point to the maintenance of the agglutinogens and agglutinins corresponding to the same blood type. These results can be expressed in four various combinations.

A. The studied blood didn't give agglutination with monoclonal antibodies that points to absence in it group agglutinogens and belonging to O(I) group. Thus Serum of the studied blood (the lower series) gives negative reaction with standard erythrocytes of O(I) group and positive – with erythrocytes of A(II) and B groups (III). It indicates existence in an blood of agglutinins α and b, i.e. confirms its belonging to O(I) group.

B. By means of monoclonal antibodies anti-A in the studied blood existence of an agglutinogen is established A. Thus Serum of the studied blood doesn't give agglutination with standard erythrocytes of O(I) and A groups (II), i.e. it doesn't contain agglutinin a, but agglutinates erythrocytes of B(III) group, i.e. in its structure there is an agglutinin b that points to belonging of an testing blood to group A (II) b.

B. By means of monoclonal antibodies anti-B in the studied blood existence of an agglutinogen of B is defined. Thus Serum of the studied blood gives negative reaction with standard erythrocytes of O(I) and B groups (III) that confirms lack of agglutinin b, and positive with erythrocytes of A(II) group that speaks about availability of agglutinin an and confirms belonging of an blood to group B (III) α.

G. By means of monoclonal antibodies anti-A and anti-B in the studied blood existence of agglutinogens A and B. In this case as control conduct research with tsoliklony anti-AB. If to it also there was an agglutination, carry out assay with a normal saline solution of chloride sodium lack of agglutination with which confirms specificity of reaction. Thus Serum of the studied blood gives negative reaction with standard erythrocytes of all three groups that specifies on lack of agglutinins in the studied blood, i.e. confirms belonging of an blood to AB (IV) group of the lake.

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